{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,20]],"date-time":"2026-04-20T13:23:40Z","timestamp":1776691420472,"version":"3.51.2"},"reference-count":38,"publisher":"American Society for Microbiology","issue":"6","license":[{"start":{"date-parts":[[2010,3,15]],"date-time":"2010-03-15T00:00:00Z","timestamp":1268611200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Appl Environ Microbiol"],"published-print":{"date-parts":[[2010,3,15]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Of the seven genetic groups, or assemblages, currently recognized in the\n            <jats:italic>Giardia duodenalis<\/jats:italic>\n            species complex, only assemblages A and B are associated with human infection, but they also infect other mammals. Recent investigations have suggested the occurrence of genetic exchanges among isolates of\n            <jats:italic>G. duodenalis<\/jats:italic>\n            , and the application of assemblage-specific PCR has shown both assemblages A and B in a significant number of human infections. In this work, three real-time quantitative (qPCR) assays were developed to target the\n            <jats:italic>G. duodenalis<\/jats:italic>\n            triose phosphate isomerase, glutamate dehydrogenase, and open reading frame C4 sequences. Primers were designed to allow the specific amplification of the DNA of assemblage A or B and to generate products distinguishable by their melting curves or, after qPCR, by their sequences, sizes, or restriction patterns. The assays showed full specificity and detected DNA from a single trophozoite (4 to 8 target copies). We applied these assays, as well as a TaqMan assay that targets the \u03b2-\n            <jats:italic>giardin<\/jats:italic>\n            gene, to genomic DNA extracted from 30 human stools and to\n            <jats:italic>Giardia<\/jats:italic>\n            cysts purified by immunomagnetic capture from the same samples. Simultaneous detection of both assemblages was observed in a large number of DNAs extracted from stools, and experiments on the cysts purified from the same samples showed that this was essentially attributable to mixed infections, as only one assemblage was detected when dilutions of cysts were tested. In a few cases, detection of both assemblages was observed even when single cysts were tested. This result, which suggests the presence of recombinants, needs to be confirmed using more accurate methods for cyst separation and enumeration. The assays described in this study can be used to detect\n            <jats:italic>Giardia<\/jats:italic>\n            cysts infectious to humans in samples from animals and in water and food.\n          <\/jats:p>","DOI":"10.1128\/aem.02305-09","type":"journal-article","created":{"date-parts":[[2010,1,16]],"date-time":"2010-01-16T02:20:30Z","timestamp":1263608430000},"page":"1895-1901","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":60,"title":["Genotyping of\n            <i>Giardia duodenalis<\/i>\n            Cysts by New Real-Time PCR Assays for Detection of Mixed Infections in Human Samples"],"prefix":"10.1128","volume":"76","author":[{"given":"Andre","family":"Almeida","sequence":"first","affiliation":[{"name":"Department of Infectious, Parasitic and Immunomediated Diseases, Istituto Superiore di Sanita\u0300, Viale Regina Elena 299, Rome, 00161, Italy"},{"name":"Centro de Imunologia e Biologia Parasita\u0301ria, Instituto Nacional de Sau\u0301de Dr. Ricardo Jorge, Rua de Sa\u0303o Lui\u0301s, 16, 4000-509 Porto, Portugal"},{"name":"Instituto de Ciencias Biome\u0301dicas de Abel Salazar, Largo Prof. Abel Salazar, 2, 4099-003 Porto, Portugal"}]},{"given":"Edoardo","family":"Pozio","sequence":"additional","affiliation":[{"name":"Department of Infectious, Parasitic and Immunomediated Diseases, Istituto Superiore di Sanita\u0300, Viale Regina Elena 299, Rome, 00161, Italy"}]},{"given":"Simone M.","family":"Caccio\u0300","sequence":"additional","affiliation":[{"name":"Department of Infectious, Parasitic and Immunomediated Diseases, Istituto Superiore di Sanita\u0300, Viale Regina Elena 299, Rome, 00161, Italy"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.14.3.447-475.2001"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.diagmicrobio.2004.06.001"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1462-5822.2001.00094.x"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.mimet.2003.11.016"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1051\/parasite\/2008153261"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.pt.2005.06.013"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.molbiopara.2008.04.006"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijpara.2008.04.008"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.exppara.2009.02.007"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cub.2007.10.020"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1000560"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.actatropica.2007.04.003"},{"key":"e_1_3_2_14_2","first-page":"259","volume":"38","year":"2007","unstructured":"Geurden, T., P. Geldhof, B. Levecke, C. Martens, D. Berkvens, S. Casaert, J. Vercruysse, and E. Claerebout. 2007. Mixed Giardia duodenalis assemblage A and E infections in calves. Int. J. Parasitol.38:259-264.","journal-title":"Int. J. Parasitol."},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.1017\/S0031182009990436"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.69.9.5178-5185.2003"},{"key":"e_1_3_2_17_2","doi-asserted-by":"publisher","DOI":"10.1086\/498169"},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.4269\/ajtmh.2007.76.713"},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0020-7519(01)00183-7"},{"key":"e_1_3_2_20_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.trstmh.2009.04.017"},{"key":"e_1_3_2_21_2","doi-asserted-by":"publisher","DOI":"10.1080\/1040-840291046713"},{"key":"e_1_3_2_22_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1550-7408.2009.00443.x"},{"key":"e_1_3_2_23_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijpara.2009.05.013"},{"key":"e_1_3_2_24_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cub.2007.11.019"},{"key":"e_1_3_2_25_2","doi-asserted-by":"publisher","DOI":"10.1017\/S0031182000064556"},{"key":"e_1_3_2_26_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.00668-07"},{"key":"e_1_3_2_27_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1143837"},{"key":"e_1_3_2_28_2","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/152.1.64"},{"key":"e_1_3_2_29_2","doi-asserted-by":"publisher","DOI":"10.1086\/517059"},{"key":"e_1_3_2_30_2","doi-asserted-by":"publisher","DOI":"10.1179\/136485908X355247"},{"key":"e_1_3_2_31_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0020-7519(01)00340-X"},{"key":"e_1_3_2_32_2","first-page":"81","volume":"27","year":"2008","unstructured":"Sahagun, J., A. Clavel, P. Goni, C. Seral, M. T. Llorente, F. J. Castillo, S. Capilla, A. Arias, and R. Gomez-Lus. 2008. Correlation between the presence of symptoms and the Giardia duodenalis genotype. Eur. J. Clin. Microbiol. Infect. Dis.27:81-83.","journal-title":"Eur. J. Clin. Microbiol. Infect. Dis."},{"key":"e_1_3_2_33_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.pt.2006.02.009"},{"key":"e_1_3_2_34_2","doi-asserted-by":"publisher","DOI":"10.1128\/EC.00138-07"},{"key":"e_1_3_2_35_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0065-308X(04)58002-8"},{"key":"e_1_3_2_36_2","unstructured":"Fighting disease fostering development. 1996"},{"key":"e_1_3_2_37_2","doi-asserted-by":"publisher","DOI":"10.1017\/S0031182007003071"},{"key":"e_1_3_2_38_2","doi-asserted-by":"publisher","DOI":"10.1051\/parasite\/200209165"},{"key":"e_1_3_2_39_2","doi-asserted-by":"publisher","DOI":"10.1007\/s10646-009-0347-9"}],"container-title":["Applied and Environmental Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/AEM.02305-09","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/AEM.02305-09","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,2,23]],"date-time":"2022-02-23T17:53:09Z","timestamp":1645638789000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/AEM.02305-09"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2010,3,15]]},"references-count":38,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2010,3,15]]}},"alternative-id":["10.1128\/AEM.02305-09"],"URL":"https:\/\/doi.org\/10.1128\/aem.02305-09","relation":{},"ISSN":["0099-2240","1098-5336"],"issn-type":[{"value":"0099-2240","type":"print"},{"value":"1098-5336","type":"electronic"}],"subject":[],"published":{"date-parts":[[2010,3,15]]},"assertion":[{"value":"2009-09-24","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2010-01-06","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2010-03-15","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}